WO2015189301A1 - Culture medium for cellular immunotherapy - Google Patents
Culture medium for cellular immunotherapy Download PDFInfo
- Publication number
- WO2015189301A1 WO2015189301A1 PCT/EP2015/062992 EP2015062992W WO2015189301A1 WO 2015189301 A1 WO2015189301 A1 WO 2015189301A1 EP 2015062992 W EP2015062992 W EP 2015062992W WO 2015189301 A1 WO2015189301 A1 WO 2015189301A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- concentration
- culture medium
- cell culture
- quality factor
- blood product
- Prior art date
Links
- 239000001963 growth medium Substances 0.000 title claims description 12
- 238000009169 immunotherapy Methods 0.000 title description 10
- 230000001413 cellular effect Effects 0.000 title description 5
- 239000010836 blood and blood product Substances 0.000 claims abstract description 109
- 229940125691 blood product Drugs 0.000 claims abstract description 109
- 239000006143 cell culture medium Substances 0.000 claims abstract description 71
- 238000000034 method Methods 0.000 claims abstract description 36
- 239000000203 mixture Substances 0.000 claims abstract description 26
- 210000004698 lymphocyte Anatomy 0.000 claims description 42
- 210000004027 cell Anatomy 0.000 claims description 39
- 210000002381 plasma Anatomy 0.000 claims description 32
- 210000004369 blood Anatomy 0.000 claims description 29
- 239000008280 blood Substances 0.000 claims description 29
- 102000004127 Cytokines Human genes 0.000 claims description 23
- 108090000695 Cytokines Proteins 0.000 claims description 23
- JYGXADMDTFJGBT-VWUMJDOOSA-N hydrocortisone Chemical compound O=C1CC[C@]2(C)[C@H]3[C@@H](O)C[C@](C)([C@@](CC4)(O)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1 JYGXADMDTFJGBT-VWUMJDOOSA-N 0.000 claims description 23
- 239000002609 medium Substances 0.000 claims description 23
- 108090000723 Insulin-Like Growth Factor I Proteins 0.000 claims description 22
- 210000002966 serum Anatomy 0.000 claims description 22
- 102000004218 Insulin-Like Growth Factor I Human genes 0.000 claims description 21
- 102100032367 C-C motif chemokine 5 Human genes 0.000 claims description 19
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 18
- 101000797762 Homo sapiens C-C motif chemokine 5 Proteins 0.000 claims description 17
- 108010002350 Interleukin-2 Proteins 0.000 claims description 17
- 102000000588 Interleukin-2 Human genes 0.000 claims description 17
- 102100030704 Interleukin-21 Human genes 0.000 claims description 17
- 108010074108 interleukin-21 Proteins 0.000 claims description 17
- 102000004889 Interleukin-6 Human genes 0.000 claims description 16
- 108090001005 Interleukin-6 Proteins 0.000 claims description 16
- 108010089417 Sex Hormone-Binding Globulin Proteins 0.000 claims description 16
- 102100030758 Sex hormone-binding globulin Human genes 0.000 claims description 16
- 102000003812 Interleukin-15 Human genes 0.000 claims description 15
- 108090000172 Interleukin-15 Proteins 0.000 claims description 15
- 102000004169 proteins and genes Human genes 0.000 claims description 15
- 108090000623 proteins and genes Proteins 0.000 claims description 15
- VOXZDWNPVJITMN-ZBRFXRBCSA-N 17β-estradiol Chemical compound OC1=CC=C2[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CCC2=C1 VOXZDWNPVJITMN-ZBRFXRBCSA-N 0.000 claims description 14
- 229960005309 estradiol Drugs 0.000 claims description 14
- 229930182833 estradiol Natural products 0.000 claims description 14
- 102000000589 Interleukin-1 Human genes 0.000 claims description 13
- 108010002352 Interleukin-1 Proteins 0.000 claims description 13
- 239000000427 antigen Substances 0.000 claims description 12
- 102000036639 antigens Human genes 0.000 claims description 12
- 108091007433 antigens Proteins 0.000 claims description 12
- 229960000890 hydrocortisone Drugs 0.000 claims description 12
- 150000002632 lipids Chemical class 0.000 claims description 12
- 108010017080 Granulocyte Colony-Stimulating Factor Proteins 0.000 claims description 10
- 102000004269 Granulocyte Colony-Stimulating Factor Human genes 0.000 claims description 10
- 108010017213 Granulocyte-Macrophage Colony-Stimulating Factor Proteins 0.000 claims description 10
- 102100039620 Granulocyte-macrophage colony-stimulating factor Human genes 0.000 claims description 10
- DDRJAANPRJIHGJ-UHFFFAOYSA-N creatinine Chemical compound CN1CC(=O)NC1=N DDRJAANPRJIHGJ-UHFFFAOYSA-N 0.000 claims description 10
- 238000012258 culturing Methods 0.000 claims description 10
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 claims description 10
- -1 CXCL-10 Proteins 0.000 claims description 9
- 102000003816 Interleukin-13 Human genes 0.000 claims description 9
- 108090000176 Interleukin-13 Proteins 0.000 claims description 9
- 102000004388 Interleukin-4 Human genes 0.000 claims description 9
- 108090000978 Interleukin-4 Proteins 0.000 claims description 9
- 108010002616 Interleukin-5 Proteins 0.000 claims description 9
- 102000000743 Interleukin-5 Human genes 0.000 claims description 9
- 108010002586 Interleukin-7 Proteins 0.000 claims description 9
- 102000000704 Interleukin-7 Human genes 0.000 claims description 9
- 102000004890 Interleukin-8 Human genes 0.000 claims description 9
- 108090001007 Interleukin-8 Proteins 0.000 claims description 9
- 108010002335 Interleukin-9 Proteins 0.000 claims description 9
- 102000000585 Interleukin-9 Human genes 0.000 claims description 9
- CVSVTCORWBXHQV-UHFFFAOYSA-N anhydrous creatine Natural products NC(=[NH2+])N(C)CC([O-])=O CVSVTCORWBXHQV-UHFFFAOYSA-N 0.000 claims description 9
- 229910052757 nitrogen Inorganic materials 0.000 claims description 9
- 102100021943 C-C motif chemokine 2 Human genes 0.000 claims description 8
- 102100023688 Eotaxin Human genes 0.000 claims description 8
- 101710139422 Eotaxin Proteins 0.000 claims description 8
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 8
- 101710091439 Major capsid protein 1 Proteins 0.000 claims description 8
- 239000008103 glucose Substances 0.000 claims description 8
- VBEQCZHXXJYVRD-GACYYNSASA-N uroanthelone Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CS)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)C(C)C)[C@@H](C)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CS)NC(=O)[C@@H](NC(=O)CNC(=O)CNC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CS)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CS)NC(=O)CNC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O)C(C)C)[C@@H](C)CC)C1=CC=C(O)C=C1 VBEQCZHXXJYVRD-GACYYNSASA-N 0.000 claims description 8
- 102000003814 Interleukin-10 Human genes 0.000 claims description 7
- 108090000174 Interleukin-10 Proteins 0.000 claims description 7
- 102400001368 Epidermal growth factor Human genes 0.000 claims description 6
- 101800003838 Epidermal growth factor Proteins 0.000 claims description 6
- 101000815628 Homo sapiens Regulatory-associated protein of mTOR Proteins 0.000 claims description 6
- 101000652747 Homo sapiens Target of rapamycin complex 2 subunit MAPKAP1 Proteins 0.000 claims description 6
- 101000648491 Homo sapiens Transportin-1 Proteins 0.000 claims description 6
- 102100028748 Transportin-1 Human genes 0.000 claims description 6
- 108060008682 Tumor Necrosis Factor Proteins 0.000 claims description 6
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims description 6
- 150000001413 amino acids Chemical class 0.000 claims description 6
- 229940116977 epidermal growth factor Drugs 0.000 claims description 6
- 102000003390 tumor necrosis factor Human genes 0.000 claims description 6
- 102000004877 Insulin Human genes 0.000 claims description 5
- 108090001061 Insulin Proteins 0.000 claims description 5
- 101710151805 Mitochondrial intermediate peptidase 1 Proteins 0.000 claims description 5
- LCTONWCANYUPML-UHFFFAOYSA-M Pyruvate Chemical compound CC(=O)C([O-])=O LCTONWCANYUPML-UHFFFAOYSA-M 0.000 claims description 5
- PNNCWTXUWKENPE-UHFFFAOYSA-N [N].NC(N)=O Chemical compound [N].NC(N)=O PNNCWTXUWKENPE-UHFFFAOYSA-N 0.000 claims description 5
- 239000004202 carbamide Substances 0.000 claims description 5
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 claims description 5
- 229960003624 creatine Drugs 0.000 claims description 5
- 239000006046 creatine Substances 0.000 claims description 5
- 229940109239 creatinine Drugs 0.000 claims description 5
- 235000014113 dietary fatty acids Nutrition 0.000 claims description 5
- 229930195729 fatty acid Natural products 0.000 claims description 5
- 239000000194 fatty acid Substances 0.000 claims description 5
- 150000004665 fatty acids Chemical class 0.000 claims description 5
- 229940125396 insulin Drugs 0.000 claims description 5
- 150000002576 ketones Chemical class 0.000 claims description 5
- 235000005985 organic acids Nutrition 0.000 claims description 5
- 150000003904 phospholipids Chemical class 0.000 claims description 5
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 claims description 4
- 150000003626 triacylglycerols Chemical class 0.000 claims description 3
- 108010055166 Chemokine CCL5 Proteins 0.000 claims description 2
- 101600111816 Homo sapiens Sex hormone-binding globulin (isoform 1) Proteins 0.000 claims description 2
- 102000008070 Interferon-gamma Human genes 0.000 claims description 2
- 108010074328 Interferon-gamma Proteins 0.000 claims description 2
- 102300044179 Sex hormone-binding globulin isoform 1 Human genes 0.000 claims description 2
- 230000029036 donor selection Effects 0.000 claims description 2
- 229960003130 interferon gamma Drugs 0.000 claims description 2
- 229940100601 interleukin-6 Drugs 0.000 claims 8
- 108010065805 Interleukin-12 Proteins 0.000 claims 5
- 102000013462 Interleukin-12 Human genes 0.000 claims 5
- 102000013691 Interleukin-17 Human genes 0.000 claims 5
- 108050003558 Interleukin-17 Proteins 0.000 claims 5
- 102000010780 Platelet-Derived Growth Factor Human genes 0.000 claims 5
- 108010038512 Platelet-Derived Growth Factor Proteins 0.000 claims 5
- 229940028885 interleukin-4 Drugs 0.000 claims 5
- 229940100602 interleukin-5 Drugs 0.000 claims 5
- 229940100994 interleukin-7 Drugs 0.000 claims 5
- 229940096397 interleukin-8 Drugs 0.000 claims 5
- XKTZWUACRZHVAN-VADRZIEHSA-N interleukin-8 Chemical compound C([C@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@@H](NC(C)=O)CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CCSC)C(=O)N1[C@H](CCC1)C(=O)N1[C@H](CCC1)C(=O)N[C@@H](C)C(=O)N[C@H](CC(O)=O)C(=O)N[C@H](CCC(O)=O)C(=O)N[C@H](CC(O)=O)C(=O)N[C@H](CC=1C=CC(O)=CC=1)C(=O)N[C@H](CO)C(=O)N1[C@H](CCC1)C(N)=O)C1=CC=CC=C1 XKTZWUACRZHVAN-VADRZIEHSA-N 0.000 claims 5
- 229940118526 interleukin-9 Drugs 0.000 claims 5
- 102100025248 C-X-C motif chemokine 10 Human genes 0.000 claims 1
- 101710098275 C-X-C motif chemokine 10 Proteins 0.000 claims 1
- 102000019223 Interleukin-1 receptor Human genes 0.000 claims 1
- 108050006617 Interleukin-1 receptor Proteins 0.000 claims 1
- 102000013275 Somatomedins Human genes 0.000 claims 1
- 229940076144 interleukin-10 Drugs 0.000 claims 1
- 239000000018 receptor agonist Substances 0.000 claims 1
- 229940044601 receptor agonist Drugs 0.000 claims 1
- 239000000306 component Substances 0.000 description 14
- 210000000987 immune system Anatomy 0.000 description 9
- 238000012360 testing method Methods 0.000 description 9
- 210000002865 immune cell Anatomy 0.000 description 7
- 244000052769 pathogen Species 0.000 description 7
- 210000001744 T-lymphocyte Anatomy 0.000 description 6
- 238000004848 nephelometry Methods 0.000 description 5
- 230000001717 pathogenic effect Effects 0.000 description 5
- 210000000130 stem cell Anatomy 0.000 description 5
- 108010039209 Blood Coagulation Factors Proteins 0.000 description 4
- 102000015081 Blood Coagulation Factors Human genes 0.000 description 4
- 206010028980 Neoplasm Diseases 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 239000003114 blood coagulation factor Substances 0.000 description 4
- 230000007423 decrease Effects 0.000 description 4
- 150000007524 organic acids Chemical class 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 230000035755 proliferation Effects 0.000 description 4
- 230000035899 viability Effects 0.000 description 4
- 102100025570 Cancer/testis antigen 1 Human genes 0.000 description 3
- 238000002965 ELISA Methods 0.000 description 3
- 101000856237 Homo sapiens Cancer/testis antigen 1 Proteins 0.000 description 3
- 241000124008 Mammalia Species 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 238000004113 cell culture Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000003102 growth factor Substances 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 241000711549 Hepacivirus C Species 0.000 description 2
- 241000724675 Hepatitis E virus Species 0.000 description 2
- 101000599951 Homo sapiens Insulin-like growth factor I Proteins 0.000 description 2
- 102100037852 Insulin-like growth factor I Human genes 0.000 description 2
- 238000002617 apheresis Methods 0.000 description 2
- 210000000601 blood cell Anatomy 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 230000004069 differentiation Effects 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 150000004676 glycans Chemical class 0.000 description 2
- 229940088597 hormone Drugs 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 229960005419 nitrogen Drugs 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 238000002616 plasmapheresis Methods 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 2
- 238000004879 turbidimetry Methods 0.000 description 2
- 229940045136 urea Drugs 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 description 2
- QGZKDVFQNNGYKY-OUBTZVSYSA-N Ammonia-15N Chemical compound [15NH3] QGZKDVFQNNGYKY-OUBTZVSYSA-N 0.000 description 1
- 208000023275 Autoimmune disease Diseases 0.000 description 1
- 102100036301 C-C chemokine receptor type 7 Human genes 0.000 description 1
- 108010012236 Chemokines Proteins 0.000 description 1
- 102000019034 Chemokines Human genes 0.000 description 1
- 102100023804 Coagulation factor VII Human genes 0.000 description 1
- 208000035473 Communicable disease Diseases 0.000 description 1
- 108010023321 Factor VII Proteins 0.000 description 1
- 108010049003 Fibrinogen Proteins 0.000 description 1
- 102000008946 Fibrinogen Human genes 0.000 description 1
- 208000031886 HIV Infections Diseases 0.000 description 1
- 241000700721 Hepatitis B virus Species 0.000 description 1
- 101000716065 Homo sapiens C-C chemokine receptor type 7 Proteins 0.000 description 1
- 101001055157 Homo sapiens Interleukin-15 Proteins 0.000 description 1
- 101001002657 Homo sapiens Interleukin-2 Proteins 0.000 description 1
- 101001010621 Homo sapiens Interleukin-21 Proteins 0.000 description 1
- 241000701085 Human alphaherpesvirus 3 Species 0.000 description 1
- 241000725303 Human immunodeficiency virus Species 0.000 description 1
- 241000713772 Human immunodeficiency virus 1 Species 0.000 description 1
- 241000713340 Human immunodeficiency virus 2 Species 0.000 description 1
- 102000014150 Interferons Human genes 0.000 description 1
- 108010050904 Interferons Proteins 0.000 description 1
- 102000015696 Interleukins Human genes 0.000 description 1
- 108010063738 Interleukins Proteins 0.000 description 1
- 102000008072 Lymphokines Human genes 0.000 description 1
- 108010074338 Lymphokines Proteins 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 108010094028 Prothrombin Proteins 0.000 description 1
- 102100027378 Prothrombin Human genes 0.000 description 1
- 241000125945 Protoparvovirus Species 0.000 description 1
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 1
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 1
- 229920002125 Sokalan® Polymers 0.000 description 1
- PZBFGYYEXUXCOF-UHFFFAOYSA-N TCEP Chemical compound OC(=O)CCP(CCC(O)=O)CCC(O)=O PZBFGYYEXUXCOF-UHFFFAOYSA-N 0.000 description 1
- 241000223109 Trypanosoma cruzi Species 0.000 description 1
- 108050007852 Tumour necrosis factor Proteins 0.000 description 1
- 102000018594 Tumour necrosis factor Human genes 0.000 description 1
- 230000033289 adaptive immune response Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 229940127219 anticoagulant drug Drugs 0.000 description 1
- 230000000890 antigenic effect Effects 0.000 description 1
- 230000008267 autocrine signaling Effects 0.000 description 1
- 210000003719 b-lymphocyte Anatomy 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 239000012503 blood component Substances 0.000 description 1
- 210000001772 blood platelet Anatomy 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 230000022131 cell cycle Effects 0.000 description 1
- 230000005754 cellular signaling Effects 0.000 description 1
- 239000000084 colloidal system Substances 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 210000004443 dendritic cell Anatomy 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000003792 electrolyte Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 229940012413 factor vii Drugs 0.000 description 1
- 229940012952 fibrinogen Drugs 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 210000003714 granulocyte Anatomy 0.000 description 1
- 239000000122 growth hormone Substances 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 102000056003 human IL15 Human genes 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 229940047124 interferons Drugs 0.000 description 1
- 229940047122 interleukins Drugs 0.000 description 1
- 210000004962 mammalian cell Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 210000001616 monocyte Anatomy 0.000 description 1
- 210000000822 natural killer cell Anatomy 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 210000005259 peripheral blood Anatomy 0.000 description 1
- 239000011886 peripheral blood Substances 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 229940039716 prothrombin Drugs 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 238000009877 rendering Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000003118 sandwich ELISA Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000002764 solid phase assay Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 230000002747 voluntary effect Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/0018—Culture media for cell or tissue culture
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0634—Cells from the blood or the immune system
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0634—Cells from the blood or the immune system
- C12N5/0636—T lymphocytes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0634—Cells from the blood or the immune system
- C12N5/0642—Granulocytes, e.g. basopils, eosinophils, neutrophils, mast cells
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0634—Cells from the blood or the immune system
- C12N5/0645—Macrophages, e.g. Kuepfer cells in the liver; Monocytes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2500/00—Specific components of cell culture medium
- C12N2500/30—Organic components
- C12N2500/32—Amino acids
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2500/00—Specific components of cell culture medium
- C12N2500/30—Organic components
- C12N2500/34—Sugars
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2500/00—Specific components of cell culture medium
- C12N2500/30—Organic components
- C12N2500/36—Lipids
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/105—Insulin-like growth factors [IGF]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/11—Epidermal growth factor [EGF]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/115—Basic fibroblast growth factor (bFGF, FGF-2)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/135—Platelet-derived growth factor [PDGF]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/20—Cytokines; Chemokines
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/20—Cytokines; Chemokines
- C12N2501/23—Interleukins [IL]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/20—Cytokines; Chemokines
- C12N2501/23—Interleukins [IL]
- C12N2501/2302—Interleukin-2 (IL-2)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/20—Cytokines; Chemokines
- C12N2501/23—Interleukins [IL]
- C12N2501/2306—Interleukin-6 (IL-6)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/20—Cytokines; Chemokines
- C12N2501/23—Interleukins [IL]
- C12N2501/2315—Interleukin-15 (IL-15)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/20—Cytokines; Chemokines
- C12N2501/23—Interleukins [IL]
- C12N2501/2321—Interleukin-21 (IL-21)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/20—Cytokines; Chemokines
- C12N2501/25—Tumour necrosing factors [TNF]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/30—Hormones
- C12N2501/33—Insulin
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/30—Hormones
- C12N2501/375—Thyroid stimulating hormone [TSH]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/30—Hormones
- C12N2501/38—Hormones with nuclear receptors
- C12N2501/39—Steroid hormones
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/30—Hormones
- C12N2501/38—Hormones with nuclear receptors
- C12N2501/39—Steroid hormones
- C12N2501/392—Sexual steroids
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/998—Proteins not provided for elsewhere
Definitions
- the present invention relates to cell culture media with high quality standards, in particular for growth and expansion immune cells of and a method of preparation.
- Active immunotherapies stimulate the patient's immune system with the intent of promoting an antigen specific anti-tumor effect using the body's own immune cells.
- the immune cells are cultured in-vitro to increase the number of cells and influence the differentiation.
- Cell culture media formulations have been well documented in the literature and a number of media are commercially available.
- Cell culture media generally comprise an appropriate source of energy and compounds which regulate the cell cycle.
- a typical culture medium is composed of a complement of amino acids, vitamins, inorganic salts, glucose, and serum as a source of growth factors, hormones, and attachment factors. In addition to nutrients, the medium also helps maintain pH and osmolality.
- Cell culture media formulations have been well documented in the literature and a number of media are commercially available.
- the cell culture medium is derived from serum or plasma as these provide an environment for the immune cells that is similar to the natural environment of the body.
- the present invention is inter alia based on the finding that the quality of the used cell culture medium has a major influence on the outcome of an expansion process of the lymphocytes for cellular immunotherapy.
- the present inventors have defined a variety of quality factors that determine the quality of cell culture medium with respect to lymphocyte expansion.
- the inventors were able to determine quality factors such as estradiol, Cortisol, IGF-1 , insulin and SHBG have to be present in certain predefined concentrations.
- a cell culture medium fulfilling the criteria determined for the individual quality factors provides an improved outcome of the culturing of mammalian cells, in particular cells of the immune system.
- the present invention provides a method for preparing a cell culture medium comprising a mixture of blood products from two or more donors, comprising the steps of:
- the invention further comprises a cell culture medium, comprising based on the volume of the cell culture medium:
- IL-10 in a concentration of below 200 pg/ml for;
- IL-13 in a concentration of below 50 pg /ml for;
- IL-1 b IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL12p70, IL-15, IL-17a, IL-21 , basic FGF, EGF, IFNy, GCSF, GM-CSF, MCP1 , MIP1 a, MIP 1 b, PDGF, IL- 1 RA, and/or TNFa in a concentration of below 20 pg/ml;
- IGF-1 in a concentration of at least 80 ⁇ 9/ ⁇ , preferably at least 120 ⁇ 9/ ⁇ , more preferably at least 140 g/l;
- the invention provides the use of the cell culture medium for culturing cells.
- the inventors have found that for cell culture immunotherapy much higher quality standards with regard to the cell culture medium have to be met. As shown in the examples, differences in the respective quality factors lead to significant changes in the outcome of lymphocyte suspension. In particular, with a medium in which the quality factors are in the predetermined ranges, growth, proliferation or expansion of immune cells lead to an increased proliferation rate, viability and activity of the cells. Identification of these quality factors allows the determination of an improved method for preparing a cell culture medium, in particular for cell populations for immunotherapy.
- the present invention provides a method for preparing a cell culture medium comprising a mixture of blood products from two or more donors, comprising the steps of:
- the method according to the invention provides a cell culture medium with high quality that in combination with factors for lymphocyte expansion allows a strong proliferation of the cells and a high activity of the expanded lymphocytes. Moreover, the method ensures the provision of a consistent quality cell culture media for facilitating the in vitro expansion of lymphocytes such as T-cells.
- cell culture medium or “growth medium” is a liquid or gel designed to support the growth of animal or plant cells. Growth media can vary in pH, glucose concentration, growth factors and the presence of other nutrients.
- blood product based media contain at least one blood product.
- synthetic medium is a chemically define medium. Such a medium does not contain any nature derived broth such as blood products. Instead, all components are added to the medium in defined concentrations.
- blood product refers to whole blood of a mammal or subsets of whole blood, in particular plasma, serum or further subsets of plasma and serum.
- processed blood product include plasma, serum and subset thereof.
- Plasma refers to the blood plasma of mammals. It is a pale white sometimes yellow liquid component of blood that normally holds the blood cells in whole blood in suspension. Plasma is mostly water and contains dissolved proteins, glucose, clotting factors, electrolytes, hormones and carbon dioxide. Blood plasma can be prepared from blood, for example by centrifugation of fresh blood containing an anticoagulant in which the blood cells fall to the bottom of the centrifugation tube. The supernatant is then the blood plasma.
- Standard as used herein is a blood plasma without coagulation factors.
- Coagulation factor as used herein also referred to as “clotting factor” includes a variety of proteins, such as fibrinogen, prothrombin or factor VII.
- Subset of whole blood refers to a solution or a suspension derived from whole blood comprising a part of the components of whole blood. Plasma and serum are subsets of whole blood.
- Donor refers to a mammal, in particular a human, from which a blood product is obtained.
- the donation maybe whole blood (WB) or specific components of whole blood retrieved by apheresis.
- Plasmapheresis as used herein is a medical technology in which blood of a donor is passed through an apparatus that separates out one particular constituent and returns the remainder to the circulation. For example, plasmapheresis leads to a collection of only blood plasma; all other components are returned to the donor.
- obtaining a blood product refers to the act of retrieving blood from a donor, in particular by taking the blood directly from the vein of the donor.
- Cytokines are a broad and loose category of small proteins ( ⁇ 5- 20 kDa) that are important in cell signaling. They are released by cells and affect the behavior of other cells. Cytokines can also be involved in autocrine signaling. Cytokines include chemokines, interferons, interleukins, lymphokines, tumour necrosis factor and growth factors.
- Chronic relevant lymphocytes are also referred to as antigen-edited lymphocytes.
- the term clinically relevant is also used for subgroups of lymphocytes.
- Particularly preferred clinically relevant lymphocytes are clinically relevant T-cells or antigen-edited T-cells.
- Clinically relevant antigens are antigens involved in a disease. Accordingly, clinically relevant antigens can be tumor-associated antigens TAA, pathogen associated antigens (PAA) or autoantigens. Tumor- reactive lymphocytes are specific for and interact with TAAs. Infectious disease reactive lymphocytes are specific for and interact with PAAs and autoimmune disease reactive lymphocytes are specific for and interact with autoantigens.
- an "antigen" is any structural substance which serves as a target for the receptors of an adaptive immune response, TCR or antibody, respectively.
- Antigens are in particular proteins, polysaccharides, lipids and substructures thereof such as peptides. Lipids and nucleic acids are in particular antigenic when combined with proteins or polysaccharides.
- a quality factor can be any component that may be found in the blood and the concentration of which has an impact on the growth and properties of a highly sensitive cell line such as cells of the immune system and stem cells.
- Cells of the immune system include, but are not limited to, B-cells, T-cells, NK cells, monocytes, dendritic cells, granulocytes and platelets.
- Blood derived media have the disadvantage that there is a variance of the ingredients due to fluctuations from one donor to the next.
- pathogen related factors are factors that identify the presence of pathogens in the blood. Commonly tested pathogens are for example anti- Trypanosoma cruzi, hepatitis B virus (HBV), hepatitis C virus (HCV), parvo virus, varicella-zoster virus, hepatitis E virus (HEV) and human immunodeficiency viruses types 1 and 2 (HIV-1 , HIV-2).
- HBV hepatitis B virus
- HCV hepatitis C virus
- HEV hepatitis E virus
- HAV-1 human immunodeficiency viruses types 1 and 2
- HIV-1 HIV-1 , HIV-2
- a quality factor is not a pathogen related factor.
- Human blood donors comprise voluntary donors and professional monitored donors. Both donors have to be healthy individuals. Furthermore, human blood donors have to fulfill certain criteria such as age, body weight and diet. Professional monitored blood donors are entered into a registry. In the registry a variety of data about the donor and the provided blood samples is recorded.
- Providing a first blood product includes in particular the transfer of the blood product into a blood bag kit, bag, container, vessel or multiwell plate.
- a first blood product from a first donor is transferred in a blood bag kit.
- a first blood product from the first donor is transferred into a multiwell plate.
- a multiwell plate is in particular a 24-well plate, a 48-well plate, a 96-well plate or a 192-well plate.
- a multiwell plate is a plate with a multiplicity of wells. The number of wells is not limited.
- the quality factor is measured by a method selected from nephelometry, turbidimetry and ELISA.
- a nephelometer is an instrument for measuring the concentration of suspended particles in a liquid or gas colloid.
- Nephelometry and turbidometry are defined in Euronorm EN 27027 (identical to the German DIN-Norm the ISO Norm 7027.
- a nephelometer measures suspended particulates by employing a light beam and a light detector set to one side of the source beam.
- antibodies specific for the quality factors are added. Interaction of the antibodies with the quality factors then leads to a participation causing a side scattering of the light. Thus, either the increase or decrease can be qualified.
- nephelometry the increase of side-scattered light is measured.
- turbidimetry the decrease of forward-scattered light is measured.
- NTU nephelometric turbidity units
- Enzyme-linked immunosorbent assay is a solid-phase assay for determining the presence and quality of antigens, in particular a sandwich ELISA can be used. The presence of pathogen related factors is in particular determined by PCR-based methods.
- Comparing the measured concentration of a quality factor to concentration range predefined for the quality factor means that a read out obtained from the measurement for a specific quality factor is taken and it is checked whether the value of the quality factor is in the range of the predetermined range.
- the testing may be done automatically by the testing apparatus or an apparatus connected to the testing apparatus. Alternatively, the comparison can be performed manually by a person carrying out measurement.
- a preferred quality factor according to the invention is a cytokine or a number of cytokines.
- the quality factor can be a group of cytokines.
- the quality factor can be all cytokines.
- a quality factor is a cytokine.
- other components have been found to strongly influence the quality of the expansion medium with regard to the ability to grow, in particular expand, cells of the immune system.
- a medium with a quality factor outside of the predefined range, i.e. non-selected medium can still be useful for growing cells for recombinant protein expression.
- Further quality factors that have been identified are estradiol, Cortisol, sex hormone-binding globulin (SHBG), insulin and insulin-like growth factor 1 (IGF-1 ).
- At least one quality factor is selected from the group consisting of estradiol, Cortisol, SHBG, insulin and IGF-1.
- CCL5 also known as RANTES.
- the predefined range for CCL5 is below 5, in particular below 3 ng/ml.
- concentration values are as defined by Nephelometrie according to ISO 7027. Accordingly, the upper threshold is 5 ng/ml, in particular 3 ng/ml; all concentrations defined for the quality factors are based on the volume of the blood product.
- a further quality factor is the cytokine Eotaxin.
- the predefined range of Eotaxin is below 800 pg/ml, in particular 500 pg/ml.
- Another quality factor is PDGF-88.
- the predefined range of PDGF-88 is below 150 pg/ml, in particular below 100 pg/ml or.
- a further quality factor is CXCL-10.
- the predefined range of CXCL-10 is below 150 pg/ml, in particular below 100 pg/ml.
- a further quality factor is the cytokine IL- 10.
- the predefined range for IL-10 is below 200 pg/ml.
- Another quality factor is the cytokine IL-13.
- the predefined range of IL-13 is below 80 pg/ml, in particular below 50 pg/ml.
- a further quality factor is I L-1 b.
- the predefined range of I L-1 b is below 30 pg/ml , in particular below 20 pg/ml.
- a further quality factor is IL-2.
- Another quality factor is IL-4.
- IL-5 is another quality factor.
- IL-7 is another quality factor.
- IL-8 is another quality factor.
- a further quality factor is IL-9.
- IL-12p70 is a quality factor.
- Another quality factor is I L-15.
- One quality factor according to the invention is IL-17a.
- IL-21 is another quality factor.
- a further quality factor is basic FGF.
- Epidermal growth factor is a further quality factor.
- Another quality factor is interferon gamma (IFNy).
- Granulocyte-colony stimulating factor (GCSF) is another quality factor according to the invention.
- Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a further quality factor according to the invention.
- MCP1 was determined as a quality factor.
- Another quality factor determined is MIP1a.
- a further quality factor is PDGF.
- IL-1 RA is another quality factor and the tumor necrosis factor a (TNFa) is another quality factor. All these factors are known cytokines that may be present in the human blood.
- IL-1 b IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL12p70, IL-15, IL-17a, IL-21 , basic FGF, EGF, IFNy, GCSF, GM-CSF, MCP1 , MIP1 a, MIP 1 b, PDGF, 111 RA, and TNFa is below 30 pg/ml , in particular below 20 pg/ml.
- the quality factor is estradiol.
- Estradiol should be in blood product with a concentration of at least 65 pmol/l. A preferred concentration for estradiol is at least 75 pmol/l. More preferably, the concentration of estradiol is at least 85 pmol/l.
- the quality factor is Cortisol.
- the predefined range for Cortisol is at least 190 nmol/l, preferably at least 210 nmol/l, more preferably at least 220 nmol/l.
- the quality factor is insulin-like growth factor 1 (IGF-1 ).
- IGF-1 insulin-like growth factor 1
- the predefined range for IGF-1 is at least 100 g/l, preferably at least 130 g/l, more preferably at least 140 Mg/I.
- cell culture medium according to the invention should fulfill. In particular, standard culture factors being present in predefined ranges.
- the method further comprises measuring standard culture factors and comparing them to predefined ranges
- standard culture factors are: proteins, glucose, non-protein nitrogen, urea nitrogen, amino acid nitrogen, creatinine, creatine, urea, total lipids, triglyceride, cholesterine, lipids, esterified components, phospholipids, fatty acids, organic acids, pyruvate, citrate, ketones.
- the predefined range based on the total volume of the cell culture medium is for proteins 60.080.0 g/l; for glucose 4.5 to 5.5 mmol/l, for non-protein nitrogen 15 to 30 mmol/l, for urea nitrogen 3.5 to 7.0 mmol/l, for amino acid nitrogen 3.0 to 5.0 mmol/l; for creatinine 70.0 to 140.0 ⁇ / ⁇ ; for creatine 25.0 to 70.0 ⁇ / ⁇ ; for urea 3.0 to 5.0 ⁇ / ⁇ ; for total lipids 4.5 to 8.5 g/l, for triglycerides 0.6 to 2.4 mmol/l, for cholesterine 4.0 to 6.5 mmol/l, lipids in a concentration of 0.3 to 0.4 mmol/l, for esterified components 0.7 to 0.8 mmol/l, for phospholipids 2.0 to 3.0 mmol/l, for fatty acids 0.3 to 0.9 mmol/l; for organic acids 4.0 to 6.0 mmol/l,
- the steps are to be performed with more than one blood product and the selected blood product or processed blood products are combined to form a culture medium.
- From one donor only about 200 to 500 ml blood product can be obtained at once. Accordingly, for the cultivation of cells several blood products of several donors have to be combined.
- For the preparation of the cell culture medium only selected blood products are used.
- the blood product is a temporary unselected blood product.
- the blood product is a temporary unselected blood product.
- a temporary unselected blood product may become a selected blood product if it is possible to mix the temporary unselected blood product with at least one selected blood product to obtain a blended blood product with the quality factors in the defined ranges. Else the temporary unselected blood product becomes an unselected blood product.
- the present invention further provides a method for preparing a cell culture medium comprising a mixture of blood products from two or more donors comprising the steps of
- the concentration measured for the quality factor is in predefined range or else unselecting the first mixture. Also, it is preferred to measure the concentration of the quality factors directly in the blood products before adding the blood product to the mixture to form a culture medium. It is also possible to screen various mixtures of blood products and verify if these fulfill the requirements as determined by the quality factors.
- the blood product can be converted into a processed blood product after selection and before addition to the final product, i.e. the cell culture medium.
- Processing the blood product means rendering one type of blood product into another type of blood product.
- the blood product measured and selected is whole blood which is then further converted into plasma.
- the measured and selected blood product is whole blood and the processed blood product is serum.
- the selected blood product is plasma and the processed blood product is serum.
- the selected blood product is plasma and the processed blood product is subset of serum.
- the selected blood product can be serum and the processed blood product is a subset of serum.
- the blood product that is provided is plasma.
- Plasma is frequently used for cultivation of for example cells of the immune system.
- plasma can be easily retrieved by plasmapheresis from the donor.
- the processed blood product is plasma.
- Using plasma has the advantage that no further process step is necessary. Thus, saving the processing step.
- two or more quality factors are tested, for example 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, 20 quality factors are tested. Testing more quality factors increases the chance of obtaining a final cell culture medium that indeed fulfills the higher quality standards for culturing and expanding highly active lymphocytes.
- testing a large amount of factors in one sample also leads to an increased complexity of the process.
- less than 15, less than 12, less than 10, less than 8, less than 6 quality factors are tested.
- the number of quality factors tested is in the range from 2 to 6, preferably 3 to 5, preferably 4. It was found that a certain subset of the known quality factors are sufficient to find a high likelihood that the cell culture medium is a cell medium eligible for expansion of lymphocytes or cultivation of stem cells.
- Quality factors that are shown to provide an indication that the whole sample is a high quality sample are SHBG, IGF-1 , CCL5 and IL-6.
- the quality factor is SHBG.
- IGF-1 as quality factor.
- CCL5 is a preferred quality factor.
- IL-6 is also a preferred quality factor.
- all quality factors are tested are SHBG, IGF-1 , CCL5 and IL-6.
- the test for a quality factor may also be used for the selection of suitable donors, in particular professional monitored donors. Professional monitored donors regularly donate blood product donations and are recorded in a registry.
- a blood donor can be identified as appropriate for donation of high quality blood products. Accordingly the method according to the invention provides a further selection step on the level of the donor.
- Donors that are unselected are removed from the registry for high quality blood product.
- the present invention also relates to a cell culture medium comprising, based on the volume of the cell culture medium:
- CCL5 in a concentration of below 3 ng/ml
- Eotaxin in a concentration of below 500 pg/ml
- PDGF-88 and/or CXCL-10 in a concentration of below 100 pg/ml
- IL-10 in a concentration of below 200 pg/ml for;
- IL-13 in a concentration of below 50 pg /ml for;
- IL-1 b IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL12p70, IL-15, IL-17a, IL-21 , basic FGF, EGF, IFNy, GCSF, GM-CSF, MCP1 , MIP1a, MIP 1 b, PDGF, 111 RA, and/or TNFa in a concentration of below 20 pg/ml;
- IGF-1 in a concentration of at least 100 ⁇ 9/ ⁇ , preferably at least 130 ⁇ 9/ ⁇ , more preferably at least 140 g/l; and/or
- SHBG in a concentration of below 31 nmol/l, preferably below 29 nmol/l.
- the medium provides nutritions necessary for T-cell culturing and expansion in a controlled and artificial environment enabling stable and reproducible results.
- the cell culture medium is preferably a medium obtained by the method of preparation according to the invention.
- the cell culture medium comprises at least one blood product, in particular plasma and/or serum. More preferably the blood product is serum.
- the cell culture medium comprises blood products from two or more donors. Preferably the blood products from two or more donors are selected from serum and/or plasma. More preferably, the products from two or more donors are serum.
- the cell culture medium is a synthetic medium.
- the cell culture medium according to the invention should fulfill.
- standard culture factore being present in predefined ranges.
- standard culture factors are: proteins, glucose, non-protein nitrogen, urea nitrogen, amino acid nitrogen, creatinine, creatine, urea, total lipids, triglyceride, cholesterine, lipids, esterified components, phospholipids, fatty acids, organic acids, pyruvate, citrate, ketones.
- the cell culture medium of the invention further comprises
- ketones in a concentration of 0.3 to 0.5 mmol/l.
- the cell culture medium fulfills all of the parameters mentioned above.
- the cell culture medium comprises either one of - SHBG in a concentration of below 29 nmol/l;
- IGF-1 in a concentration of at least 100 ⁇ 9/ ⁇
- IL-6 in a concentration of below 20 pg/ml
- the cell culture medium comprises
- IGF-1 in a concentration of at least 100 ⁇ 9/ ⁇
- IL-6 in a concentration of below 20 pg/ml
- the cell culture medium according to the invention may consist of a mixture of blood products.
- the cell culture medium comprises in addition to the mixture of blood products further components.
- the further components are selected from water, NaCI, PBS, DTT, TCEP, or 2-mercaptoethanol.
- the cell culture medium according to the invention is suitable for culturing a variety of different cells.
- the invention further provides the use of the cell culture medium for culturing cells.
- a major advantage is that besides the culturing of a immortal cell line also cells obtained from a patient, in particular cells of the immune system or stem cells, may be successfully cultured.
- a medium according to the invention in particular a medium obtained with a method according to the invention, much higher expansion rates of lymphocytes are found.
- “Expansion” or “clonal expansion” as used herein means production of daughter cells derived originally from a single cell. In a clonal expansion of lymphocytes, all progenies shared the same antigen specificity.
- the culture medium according to the invention in cell culture an increased total number of cells can be obtained.
- an increased number of total expanded daughter cells of lymphocytes with the same antigen specificity can be obtained.
- the cells are cells of the immune system, in particular lymphocytes.
- the lymphocytes are obtained from the patient.
- the lymphocytes are grown, proliferated and/or expanded in the cell culture medium.
- the culture medium is used in the expansion of T- cells derived from tumor.
- the expansion involves the use of a cytokine cocktail of IL-2, IL-15 and IL-21.
- the composition of IL-2, IL-15 and IL-21 also referred to as "the cytokine cocktail”.
- interleukin 2 or “IL-2” refers to human IL-2 as defined by SEQ ID NO: 1 and functional equivalents thereof.
- interleukin 15 or “IL- 15” refer to human IL-15 and functional equivalents thereof.
- interleukin 21 " or “IL-21” refer to human IL-21 and functional equivalents thereof.
- the cell culture medium according to the invention preferably a medium obtained by the method according to the invention, is in particular suitable for the expansion of lymphocytes in combination with a cytokine mixture of IL-2, IL-15 and/or IL-21.
- the cell culture medium according to the invention provides an increased growth rate, an increased total number of expanded cells but also an improved profile of the cells.
- the maturation and differentiation defined by the CD45RA CCR7 phenotype is improved.
- an expansion of lymphocytes with a mixture of cytokines IL-2, IL-15 and IL-21 leads to an increased viability of clinically relevant lymphocytes, in particular T-cells.
- the increased growth and expansion rate decreases the time from obtaining the lymphocyte sample from the patient to the time of reintroducing the standard lymphocytes into the patient.
- the increased viability of the (clinically relevant) lymphocytes leads to an increased therapeutical effect of the active immunotherapy product.
- Example 1 Expansion of lymphocytes in different cell culture media.
- Aphaeresis was performed on a healthy donor primed with NY-ESO-1 . After separation of the blood components, the product containing the leucocytes is separated from the rest.
- Cells were suspended in medium M 1 with 1000 U/ml IL-2, 10 ng/ml IL-15 and 10 ng/ml IL-21 .
- the medium in addition contained 10 ⁇ NY-ESO-1 peptide.
- the cells expanded well reaching a concentration of 10 6 /ml after 4 days.
- lymphocytes were expanded in medium M2 according to the same protocol. No significant expansion of lymphocytes was detected.
- the plasma samples P1 to P4 are unselected.
- Plasma sample P5 is selected for lymphocyte culture.
- Lymphocytes obtained from peripheral blood of a healthy donor primed with NY-ESO-1 were cultured according to the above described protocol. Expansion in cell culture medium from P5 was comparable to the expansion in medium M1 (see Example 1 ).
Landscapes
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Immunology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- General Engineering & Computer Science (AREA)
- Cell Biology (AREA)
- Hematology (AREA)
- Gastroenterology & Hepatology (AREA)
- Medicinal Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
Description
Claims
Priority Applications (9)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CA2951746A CA2951746C (en) | 2014-06-10 | 2015-06-10 | Culture medium for cellular immunotherapy |
AU2015273531A AU2015273531B2 (en) | 2014-06-10 | 2015-06-10 | Culture medium for cellular immunotherapy |
KR1020177000767A KR102573837B1 (en) | 2014-06-10 | 2015-06-10 | Culture medium for cellular immunotherapy |
RU2017100024A RU2708199C2 (en) | 2014-06-10 | 2015-06-10 | Culture medium for cell immunotherapy |
US15/317,774 US10975353B2 (en) | 2014-06-10 | 2015-06-10 | Culture medium for cellular immunotherapy |
JP2017517405A JP6806970B2 (en) | 2014-06-10 | 2015-06-10 | Culture medium for cell immunotherapy |
EP15733367.5A EP3155092B1 (en) | 2014-06-10 | 2015-06-10 | Culture medium for cellular immunotherapy |
CN201580042926.XA CN106574245A (en) | 2014-06-10 | 2015-06-10 | Culture medium for cellular immunotherapy |
IL249410A IL249410B (en) | 2014-06-10 | 2016-12-06 | Culture medium for cellular immunotherapy |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE102014211052.1 | 2014-06-10 | ||
DE102014211052 | 2014-06-10 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2015189301A1 true WO2015189301A1 (en) | 2015-12-17 |
Family
ID=53498960
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/EP2015/062992 WO2015189301A1 (en) | 2014-06-10 | 2015-06-10 | Culture medium for cellular immunotherapy |
Country Status (10)
Country | Link |
---|---|
US (1) | US10975353B2 (en) |
EP (1) | EP3155092B1 (en) |
JP (1) | JP6806970B2 (en) |
KR (1) | KR102573837B1 (en) |
CN (1) | CN106574245A (en) |
AU (1) | AU2015273531B2 (en) |
CA (1) | CA2951746C (en) |
IL (1) | IL249410B (en) |
RU (1) | RU2708199C2 (en) |
WO (1) | WO2015189301A1 (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP3154567B1 (en) * | 2014-06-11 | 2020-11-25 | polybiocept GmbH | Expansion of lymphocytes with a cytokine composition for active cellular immunotherapy |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR102631682B1 (en) * | 2021-12-02 | 2024-02-28 | (주)에스엠티바이오 | Method for selecting an appropriate donor of immune cells |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1999067365A1 (en) * | 1998-06-22 | 1999-12-29 | Medi-Cult A/S | A method for in vitro maturation of human gametes |
WO2007037682A1 (en) * | 2005-09-28 | 2007-04-05 | Ipd-Therapeutics B.V. | Methods and means for stem cell proliferation and subsequent generation and expansion of progenitor cells, as well as production of effector cells as clinical therapeutics. |
Family Cites Families (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5648350A (en) * | 1993-05-25 | 1997-07-15 | Laboratoires Besins Iscovesco | Dihydrotestosterone for use in androgenotherapy |
JP2001275662A (en) * | 2000-04-03 | 2001-10-09 | Nobuhiko Emi | Human serum and method for producing the same |
JP2003235548A (en) * | 2001-12-13 | 2003-08-26 | Japan Science & Technology Corp | Culture medium for human cell and culture method |
EP1666589B1 (en) * | 2003-08-22 | 2010-02-17 | Takara Bio Inc. | Process for producing cytotoxic lymphocytes |
CN1754574A (en) * | 2004-09-27 | 2006-04-05 | 深圳市中兴扬帆生物工程有限公司 | Use of immune cell for treating blood coloboma symptom and arrest of bone marrow |
JP4929174B2 (en) * | 2005-08-17 | 2012-05-09 | タカラバイオ株式会社 | Method for producing lymphocytes |
US8183040B2 (en) * | 2008-04-15 | 2012-05-22 | New York University | Methods for in vitro differentiation of Th-17+cells |
RU2415929C2 (en) * | 2009-06-10 | 2011-04-10 | Общество с ограниченной ответственностью Научно-производственное предприятие "ПанЭко" | Specialised medium for human amniotic fluid and fibroblast cell culture |
ES2627910T3 (en) * | 2009-12-29 | 2017-08-01 | Gamida-Cell Ltd. | Methods to enhance the proliferation and activity of natural destructive cells |
EP3875599A1 (en) * | 2010-07-23 | 2021-09-08 | Astellas Institute for Regenerative Medicine | Methods for detection of rare subpopulations of cells and highly purified compositions of cells |
-
2015
- 2015-06-10 CA CA2951746A patent/CA2951746C/en active Active
- 2015-06-10 KR KR1020177000767A patent/KR102573837B1/en active IP Right Grant
- 2015-06-10 AU AU2015273531A patent/AU2015273531B2/en active Active
- 2015-06-10 US US15/317,774 patent/US10975353B2/en active Active
- 2015-06-10 JP JP2017517405A patent/JP6806970B2/en active Active
- 2015-06-10 EP EP15733367.5A patent/EP3155092B1/en active Active
- 2015-06-10 RU RU2017100024A patent/RU2708199C2/en active
- 2015-06-10 CN CN201580042926.XA patent/CN106574245A/en active Pending
- 2015-06-10 WO PCT/EP2015/062992 patent/WO2015189301A1/en active Application Filing
-
2016
- 2016-12-06 IL IL249410A patent/IL249410B/en active IP Right Grant
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1999067365A1 (en) * | 1998-06-22 | 1999-12-29 | Medi-Cult A/S | A method for in vitro maturation of human gametes |
WO2007037682A1 (en) * | 2005-09-28 | 2007-04-05 | Ipd-Therapeutics B.V. | Methods and means for stem cell proliferation and subsequent generation and expansion of progenitor cells, as well as production of effector cells as clinical therapeutics. |
Non-Patent Citations (3)
Title |
---|
BIESECKER L G ET AL: "Interleukin-6 is a component of human umbilical cord serum and stimulates hematopoiesis in embryonic stem cells in vitro", EXPERIMENTAL HEMATOLOGY, ELSEVIER INC, US, vol. 21, no. 6, 1 June 1993 (1993-06-01), pages 774 - 778, XP009185719, ISSN: 0301-472X * |
MCNAMEE J P ET AL: "Effect of pro-inflammatory cytokines on spontaneous apoptosis in leukocyte sub-sets within a whole blood culture", CYTOKINE, ACADEMIC PRESS LTD, PHILADELPHIA, PA, US, vol. 31, no. 2, 21 July 2005 (2005-07-21), pages 161 - 167, XP004962352, ISSN: 1043-4666, DOI: 10.1016/J.CYTO.2005.05.001 * |
STEVE J. MCFAUL ET AL: "Hemoglobin stimulates the release of proinflammatory cytokines from leukocytes in whole blood", JOURNAL OF LABORATORY AND CLINICAL MEDICINE, vol. 135, no. 3, 1 March 2000 (2000-03-01), pages 263 - 269, XP055206586, ISSN: 0022-2143, DOI: 10.1067/mlc.2000.105180 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP3154567B1 (en) * | 2014-06-11 | 2020-11-25 | polybiocept GmbH | Expansion of lymphocytes with a cytokine composition for active cellular immunotherapy |
Also Published As
Publication number | Publication date |
---|---|
US20170121680A1 (en) | 2017-05-04 |
JP2017522902A (en) | 2017-08-17 |
KR20170031138A (en) | 2017-03-20 |
EP3155092B1 (en) | 2024-02-28 |
EP3155092C0 (en) | 2024-02-28 |
IL249410B (en) | 2021-01-31 |
IL249410A0 (en) | 2017-02-28 |
AU2015273531A1 (en) | 2017-01-12 |
EP3155092A1 (en) | 2017-04-19 |
KR102573837B1 (en) | 2023-09-01 |
CN106574245A (en) | 2017-04-19 |
CA2951746C (en) | 2023-08-22 |
JP6806970B2 (en) | 2021-01-06 |
AU2015273531B2 (en) | 2021-07-29 |
CA2951746A1 (en) | 2015-12-17 |
US10975353B2 (en) | 2021-04-13 |
RU2017100024A3 (en) | 2018-11-28 |
RU2708199C2 (en) | 2019-12-04 |
RU2017100024A (en) | 2018-07-18 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN110747166B (en) | In-vitro amplification culture method for peripheral blood T cells | |
CN104593324B (en) | A kind of culture medium of natural killer cells and the amplification cultivation method of natural killer cells | |
JP4275680B2 (en) | Culture methods for lymphocyte activity / proliferation | |
AU2015273531B2 (en) | Culture medium for cellular immunotherapy | |
CA3048831A1 (en) | Method for obtaining monocytes or nk cells | |
JP2021019593A (en) | Human t-cell line assay for evaluating immunologic identity of glatiramer acetate preparations | |
CN110646619A (en) | Method for detecting cell factor secreted by specific T cell in lung cancer or intestinal cancer | |
JP2011516582A (en) | Improved method of making IRX-2 | |
CN111172110B (en) | Culture method of umbilical cord blood CIK cells | |
CN115960829A (en) | Method for efficiently amplifying NK cells | |
CN110857435A (en) | Culture medium for culturing immune cells separated from cord blood and culture method thereof | |
CN102144165A (en) | Stimulated cell standards | |
CN111434674B (en) | Polypeptide composition and use thereof in cancer immunotherapy | |
JP2022533012A (en) | Systems and methods for regulating cell phenotype | |
Kozbial et al. | Automated generation of immature dendritic cells in a single-use system | |
CN111690608A (en) | Double-antibody and thymosin combined reagent for in-vitro culture of NK (natural killer) cells, kit and culture method | |
CN110964695A (en) | Cell strain and method for detecting rhIL-12 in vitro activity by proliferation thereof | |
CN112675201B (en) | Application of macrophage subgroup and regulator thereof in acute graft-versus-host disease | |
LU500984B1 (en) | Detection of Cytokines Secreted by Specific T Cells in Lung or Colorectal Cancer | |
CN118064364B (en) | Preparation method of gamma delta T cells | |
US11578132B2 (en) | Method for producing B cell population and method for producing monoclonal antibody using same | |
CN106701676B (en) | Novel NKT-like cell subsets and their use for treating immune diseases | |
Parackova et al. | Effect of sampling media and culture conditions on dendritic cell generation: ion depleted plasma negatively influences the survival of monocyte-derived dendritic cells | |
JP2023064786A (en) | Secretome detection method, and culture medium for labeling t-cell | |
CN117210404A (en) | High-purity peripheral blood autologous NK cell culture medium and in-vitro culture method |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 15733367 Country of ref document: EP Kind code of ref document: A1 |
|
WWE | Wipo information: entry into national phase |
Ref document number: 249410 Country of ref document: IL |
|
ENP | Entry into the national phase |
Ref document number: 2951746 Country of ref document: CA |
|
WWE | Wipo information: entry into national phase |
Ref document number: 15317774 Country of ref document: US |
|
ENP | Entry into the national phase |
Ref document number: 2017517405 Country of ref document: JP Kind code of ref document: A |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
ENP | Entry into the national phase |
Ref document number: 2017100024 Country of ref document: RU Kind code of ref document: A Ref document number: 20177000767 Country of ref document: KR Kind code of ref document: A |
|
REEP | Request for entry into the european phase |
Ref document number: 2015733367 Country of ref document: EP |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2015733367 Country of ref document: EP |
|
ENP | Entry into the national phase |
Ref document number: 2015273531 Country of ref document: AU Date of ref document: 20150610 Kind code of ref document: A |